Effects On Proliferation Inhibition And Expression Of PPAR-??TGF-?1 In Rat Hepatic Stellate Cells By Total Flavone From Litchi Chinensis Sonn

Objective: To observe HSC-T6 cells' proliferation suppressed and PPAR-??TGF-?1's expression affected by total flavone from Litchi Chinensis Sonn(TFL),and to discuss the anti-hepatic fibrosis' s possible mechanism of action by TFL.Methods: HSC-T6 cells were cultured in vitro and intervened by different concentration TFL lasted for 24h?48h and 72 h,then cells' suppress proliferation of the different groups were detected by CCK-8 method in different time periods.HSC-T6 cells were divided into blank control groups,Rosiglitazone groups,TFL+Rosiglitazone groups,TFL groups and TFL+GW9662 groups.After 72 h,we detect the cells' suppress proliferation in all groups by CCK-8 method and observe those morphology;and then to detect the protein content of TIMP-1?MMP-2?Collagen?and Collagen III in every groups' cell supernatants by ELISA method.;And then to detect the PPAR-?and TGF-?1 mRNA transcription by q PCR method;and then to detect the PPAR-?and TGF-?1 expression by Immunocytochemical method;At last,we make a correlation analysis of PPAR-?and TGF-?1expression by SPSS 17.Results:1)The CCK-8 results show that the inhibitory effect of TFL on the proliferation of HSC-T6 cells was increased with the prolongation of action time and the increasing of the concentration.2)The results of cell morphology observation showed that the growth of cells was inhibited, the growth of cells was decreased and the growth gap was increased.Among them,the growth inhibition of TFL+ rosiglitazone group was the most obvious,and the TFL group was more obvious than the TFL+GW9662 group.3)The experimental group showed that the cell proliferation of 72h: drug intervention after each cell proliferation was inhibited,TFL+rosiglitazone group the highest inhibition rate,TFL+GW9662 group inhibition rate lowest,inhibition rate of TFL group between TFL+ rosiglitazone group and TFL+GW9662 group.4)The ELISA results showed that compared with the blank group,the content of other groups TIMP-1,MMP-2,I and III collagen were significantly decreased(P<0.01);compared with rosiglitazone group,the content of MMP-2,TIMP-1,I and type III collagen in TFL+rosiglitazone group were significantly reduced(P<0.01),the content of TIMP-1,MMP-2,I and III collagen in TFL+GW9662 group were increased(P<0.05);compared with TFL group,the content of I type collagen,TIMP-1 and MMP-2 in the TFL+rosiglitazone group were significantly lower(P<0.01),the content of III collagen,TIMP-1,content of MMP-2 in the TFL+GW9662 group were increased(P<0.05).5)The q PCR results showed that compared with the blank group,the expression of PPAR-?mRNA in other experimental group were increased,the expression of TGF-?1 mRNA was significantly decreased(P<0.01);Compared with Rosiglitazone group,the expression of PPAR-?mRNA in TFL+Rosiglitazone group was increased,the expression of TGF-?1mRNA was decreased(P<0.01),the expression of PPAR-?mRNA in TFL+GW9662 group was decreased,the expression of TGF-?1 mRNA was increased(P<0.05).Compared with TFL group,the the expression of PPAR-?mRNA in TFL+GW9662 group was decreased,the expression of TGF-?1 mRNA in TFL+GW9662 group was increased(P<0.01).6)The Immunocytochemistry results showed that The protein was mainly located in the cytoplasm,which showed positive expression of brown staining.Compared with the blank group,the expression of PPAR-?in the other experimental groups were increased,the expression of TGF-?1were decreased(P<0.01);Compared with Rosiglitazone and TFL group,the expression of PPAR-?in TFL+Rosiglitazoneon group was increased,the expression of TGF-?1was decreased(P<0.01),the expression of PPAR-?in TFL+GW9662 group was increased,the expression of TGF-?1was decreased(P<0.01).7)The correlation analysis showed that the expression of PPAR-?and TGF-?1 was negatively correlated,and the coefficient of association were r=-7.97,r=-8.55(p<0.01).expression of PPAR-?mRNA in TFL+Rosiglitazone group was increased,Conclusion:TFL can inhibited HSC-T6 cells' proliferation,and has a dose-effect relationship.TFL can reduce the secretion of TIMP-1,MMP-2,I collagen and collagen type III in the supernatant of HSC-T6 cells.The up regulation of PPAR-? expression and the down regulation of TGF-?1 expression by TFL.The effect that TFL on anti hepatic fibrosis is associated with the fact that it can stimulate the activation of PPAR-?,and inhibited TGF-?1 signaling pathway,suppress the expression of TIMP-1 and MMP-2,reduce the excretion of I and III collagen,and reduce the intracellular deposition of ECM,all of which in turn leads to the reversing process of hepatic fibrosis.